Two-photon multiplane imaging of neural circuits
(Conference Presentation)

Weijian Yang; Jae-eun K Miller; Luis Carrillo-Reid; Eftychios Pnevmatikakis; Liam Paninski; Darcy S. Peterka; Rafael Yuste

doi:10.1117/12.2219843

26 April 2016 Two-photon multiplane imaging of neural circuits (Conference Presentation)

Weijian Yang, Jae-eun K Miller, Luis Carrillo-Reid, Eftychios Pnevmatikakis, Liam Paninski, Darcy S. Peterka, Rafael Yuste

Author Affiliations +

Proceedings Volume 9690, Clinical and Translational Neurophotonics; Neural Imaging and Sensing; and Optogenetics and Optical Manipulation; 969016 (2016) https://doi.org/10.1117/12.2219843
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

Imaging the neuronal activity throughout the brain with high temporal and spatial resolution is an important step in understanding how the brain works. Two-photon laser scanning microscopy with fluorescent calcium indicators has enabled this type of experiments in vivo. Most of these microscopes acquire images serially, with a single laser beam, limiting the overall imaging speed. To overcome this limit, multiple beamlets can be used to image in parallel multiple regions. Here, we demonstrate a novel scheme of a two-photon laser-scanning microscope that can simultaneously record neuronal activity at multiple planes of the sample with a single photomultiplier tube. A spatial light modulator is used to generate the designated multiple beamlets, and a constrained non-negative matrix factorization algorithm is used to demix the signals from multiple scanned planes. We simultaneously record neuronal activity of multiple layers of a mouse cortex at 10 fps in vivo. This novel imaging scheme provides a powerful tool for mapping the brain activity.

Conference Presentation

Citation Download Citation

Weijian Yang, Jae-eun K Miller, Luis Carrillo-Reid, Eftychios Pnevmatikakis, Liam Paninski, Darcy S. Peterka, and Rafael Yuste "Two-photon multiplane imaging of neural circuits (Conference Presentation)", Proc. SPIE 9690, Clinical and Translational Neurophotonics; Neural Imaging and Sensing; and Optogenetics and Optical Manipulation, 969016 (26 April 2016); https://doi.org/10.1117/12.2219843

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KEYWORDS

Brain

Neuroimaging

In vivo imaging

Microscopes

Two photon imaging

Calcium

Laser scanners

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